FlightlessⅠ与核质转运蛋白Importin β及核孔蛋白Nup88的相互作用

Interaction of Flightless I with Nup88 and Importin β

摘要:

Fightless I (FLII) 是一个在肿瘤中高表达的蛋白，前期研究提示FLII可能参与核质转运过程，为鉴定FLII是否与核膜结合蛋白相互作用，构建GST-FLII、GST-LRR融合蛋白重组质粒并转化至大肠杆菌 Rosetta 进行诱导表达，使用GST琼脂糖珠进行融合蛋白的纯化。通过SDS-PAGE对纯化蛋白进行验证之后，利用GST-pull down及免疫共沉淀技术证明了FLII与Importin β、Nup88蛋白的相互作用，并鉴定FLII上的LRR结构域为相互作用区域。研究结果提示FLII可能参与了Importin β的部分生物学作用，为进一步分析FLII与Importin β、Nup88的生物学功能奠定了基础。

Abstract:

High expression of Fightless I (FLII) is associated to multiple tumors. Based on our previous study that FLII might be involved in the nuclear export, we assessed the possible interaction of FLII with the nuclear envelop associating proteins Importin β and Nup88. We first constructed GST-FLII, GST-LRR recombinant plasmids and transformed them into the Rosetta strain to produce GST-FLII, GST-LRR fusion protein. After purification of these proteins, GST-pull down, as well as co-immunoprecipitation, were used to test the interaction of FLII with Importin β and Nup88. FLII interacted with Importin β and Nup88, and FLII LRR domain is responsible for these interactions. Thus, FLII may play a role in nuclear export through interaction with Importin β and Nup88.